Introduction: Staphylococcus aureus (S. aureus) is a Gram-positive cocci arranged in grape like clusters, non-motile, non-sporing, non-capsulated, that causes nosocomial infections, severe blood infections, bacteremia, food poisoning, cutaneous infections, toxic shock syndrome. Objectives: To isolate and identify S. aureus from blood samples. To perform antimicrobial susceptibility testing of S. aureus. To find prevalence of bacteremia caused due to S. aureus. Material & Methods: The study was carried out from October 2021 to March 2022, A total 1,270 samples were received in Bacteriology section from patients admitted in various ICUs & EMR of AIMSR hospital. S. aureus was identified on the basis of colony characteristics, gram staining morphology and biochemical tests. Antimicrobial susceptibility testing was done by automated method, Vitek 2 compact machine using P628AST card. Results: The prevalence of S. aureus in culture positive blood samples was 2.83%. Maximum S. aureus were obtained from CCU department and minimum from NICU & PICU. S. aureus isolation was slightly more in males 25(69.4%) than females 11 (30.6%) and was found to be same in the age group of 21-40 & 41-60 years (30.60%) as compared to other age groups. S. aureus isolates were highly sensitive to teicoplanin, vancomycin, tigecycline, doxycycline, minocycline (100%). Sensitivity towards linezolid was recorded as (96%), Tetracycline (86.1%) and Daptomycin (77.7%). However, they showed resistance to Benzylpenicillin, & Oxacillin (100%), Ciprofloxacin & Levofloxacin (83.3%), Erythromycin (69.5%), Cotrimoxazole (52.8%), Clindamycin (41.7%), Gentamicin (38.9%), Daptomycin (22.3%), Tetracycline(13.9%), Rifampicin (5.6%), Linezolid (3%). Conclusion: S. aureus bacteremia causes a health burden, particularly in low and middle income countries. S. aureus infections are a significant clinical problem in medical practice as the organism shows resistance to the commonly used first line drugs. Keywords: Staphylococcus aureus, Gram positive cocci, Bacteremia, methicillin resistant S. aureus, antimicrobial susceptibility testing.